Parkin-catalyzed Ubiquitin-Ester Transfer Is Triggered by PINK1-dependent Phosphorylation

• DOI: 10.1074/jbc.m113.467530
• Publication Date: 2013-06-11T03:33:16Z
• Authors: Masahiro Iguchi; Yuki Kujuro; Kei Okatsu; Fumika Koyano; Hidetaka Kosako; Mayumi Kimura; Norihiro Suzuki; Shinichiro Uchiyama; Keiji Tanaka; Noriyuki Matsuda
• Keywords: PINK1; Ubiquitin-Protein Ligases

PINK1 and PARKIN are causal genes for autosomal recessive familial Parkinsonism. is a mitochondrial Ser/Thr kinase, whereas Parkin functions as an E3 ubiquitin ligase. Under steady-state conditions, localizes to the cytoplasm where its activity repressed. A decrease in membrane potential triggers recruits it depolarized mitochondria ubiquitylation of substrates. The molecular basis how re-established by damage has yet be determined. Here we provide vitro biochemical evidence ubiquitin-thioester formation on Cys-431 recombinant Parkin. We also report that forms ubiquitin-ester following cells, this event essential substrate ubiquitylation. Importantly, RING2 domain acts transthiolation or acyl-transferring rather than E2-recruiting domain. Furthermore, depends phosphorylation Ser-65. phosphorylation-deficient mutation completely inhibited intermediate, mimics, such Ser Glu substitution, enabled partial intermediate irrespective Ser-65 phosphorylation. propose PINK1-dependent leads transfer reaction domain, step activation.